VEGF and Macrophage Expression in Pulp Inflammation Following Administration of a Combination of Tricalcium Silicate Cement and Galam (Melaleuca cajuputi var. cumingiana) Leaf Extract: In Vivo and In Silico Studies

Abstract

The use of natural compounds with anti-inflammatory and regenerative effects for wound healing has been found beneficial. Galam (Melaleuca cajuputi var. cumingiana) is a typical Indonesian medicinal plant with anti-inflammatory activity. This study aimed to evaluate the combined effect of galam leaf extract and tricalcium silicate cement on vascular endothelial growth factor (VEGF) and macrophage expression in pulpitis rat model. Twenty-four male Wistar rats were divided into six groups: Groups 1 and 2 - received a combination of galam leaf extract and tricalcium silicate cement for 3 and 7 days, respectively; Groups 3 and 4 - received tricalcium silicate cement alone for 3 and 7 days, respectively; Groups 5 and 6 - received 40% propylene glycol for 3 and 7 days, respectively. After the treatment periods, macrophage and VEGF expressions were assessed by histological and immunohistochemical analyses. The binding interactions of phytoconstituents of galam leaf with VEGF protein was predicted through molecular docking simulations. Results showed a significant increase in VEGF expression and a reduction in pulp inflammation in the combination groups compared to the controls. Macrophage numbers increased on day 3, indicating immune activation, and decreased by the 7th day, suggesting inflammation resolution. The compound dammarane-3,12,25-triol present in galam leaf demonstrated strong binding affinity to VEGF in silico, suggesting potential angiogenic effect. These findings indicate that the combination of galam leaf extract and tricalcium silicate cement enhanced pulp healing by reducing inflammation, regulating macrophage activity, and increasing VEGF expression in pulpitis rat model.

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